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Team:Bielefeld-Germany/Labjournal/week4
From 2012.igem.org
Contents |
Labjournal
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Week 4 (05/21 - 05/27/12)
Monday May 21st
- Team Modeling: Our aims for modeling:
- model the expression of the laccases in the organisms.
- model the aktivity of our enzyms.
- model the interesing parts of a clarification plant (the part witch are interesing for our cleaner.
- Team Bacterial Laccases: We did some restrictions to clone our laccase PCR products from Xanthomonas campestris and E. coli in pSB1C3 backbone. Therefore we transformed pSB1C3 with RFP in our competent KRX cells, picked colnies and made plasmidisolations. After that we ligated the cutted products with the cutted plasmid backbone.
Tuesday May 22nd
Team Bacterial Laccases: Transformation of the ligated plasmids in competent E. coli KRX cells.
Wednesday May 23rd
- Team Student Academy: Repetition of the transformation of BBa_J04450 and BBa_I13522 with new competent cells. For setup see here. Got intense red fluorescing colonies but no green fluorescing colonies. Made a backup of BBa_J04450. Asking for other plasmids containing GFP at the working groups of our University.
- Team Bacterial Laccases:
- After there were no colonies on our pSB1C3 + CopA (Xanthomonas campestris) plate we did the transformation with the same ligation preparation again. For the other ligation with pSB1C3 + Cueo (E. coli) we started colony PCRSs to find postive colonies. Sadly the colony PCR showed no product.
- Purification of the T. thermo laccase PCR product out of agarose gel.
Thursday May 24th
- Team Student Academy: Made a liquid culture of BBa_J04450 at 30°C. There was no fluorescence. Furthermore BBa_J23100 was transformed. Also got intense red fluorescing colonies.
Friday May 25th
Team Bacterial Laccases:
- We had to to the PCR on T. thermo laccase again, because before the amount of purified PCR product was very low.
